Spark Blue™ 574 anti-human CD69 (Flexi-Fluor™) Antibody

Pricing & Availability
Clone
FN50 (See other available formats)
Regulatory Status
RUO
Workshop
IV A91
Other Names
Very Early Activation Antigen (VEA), Activation inducer molecule (AIM)
Isotype
Mouse IgG1, κ
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Product Citations
publications
Cat # Size Price Quantity Save
285169 50 µg $394
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Description

CD69 is a 27-33 kD type II transmembrane protein also known as activation inducer molecule (AIM), very early activation antigen (VEA), and MLR3. It is a member of the C-type lectin family, expressed as a disulfide-linked homodimer. Other members of this receptor family include NKG2, NKR-P1 CD94, and Ly49. CD69 is transiently expressed on activated leukocytes including T cells, thymocytes, B cells, NK cells, neutrophils, and eosinophils. CD69 is constitutively expressed by a subset of medullary mature thymocytes, platelets, mantle B cells, and certain CD4+ T cells in germinal centers of normal lymph nodes. CD69 is involved in early events of lymphocyte, monocyte, and platelet activation, and has a functional role in redirected lysis mediated by activated NK cells.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human
Reported Reactivity
African Green, Baboon, Chimpanzee, Cynomolgus, Pigtailed Macaque, Rhesus
Antibody Type
Monoclonal
Host Species
Mouse
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography and conjugated with Spark Blue™ 574 under optimal conditions.
Concentration
0.2 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

FC

Recommended Usage

Flexi-Fluors™ are provided at a standard 0.2 mg/mL concentration. We recommend titrating this reagent to determine the optimal concentration for each application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL. We recommend testing a range of concentrations starting from 10 µg/mL.

For example, make five 1:1 serial dilutions of the 0.2 mg/mL antibody. Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/mL) to test six concentrations -- 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration.

Excitation Laser
Blue Laser (488 nm)
Application Notes

Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen tissue sections2, immunofluorescence microscopy3, and spatial biology (IBEX)8,9.

Additional Product Notes

For more information about Flexi-Fluors™, visit our Flexi-Fluor™ page and review FAQs associated with this product line.

Application References

(PubMed link indicates BioLegend citation)
  1. Knapp WB, et al. 1989. Leucocyte Typing IV. Oxford University Press. New York.
  2. Sakkas LI, et al. 1998. Clin. and Diag. Lab. Immunol. 5:430. (IHC)
  3. Kim JR, et al. 2005. BMC Immunol. 6:3. (IF)
  4. Verjans GM, et al. 2007. P. Natl. Acad. Sci. USA 104:3496.
  5. Lu H, et al. 2009. Toxicol Sci. 112:363. (FC) PubMed
  6. Thakral D, et al. 2008. J. Immunol. 180:7431. (FC) PubMed
  7. Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC)
  8. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  9. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
RRID
AB_3662509 (BioLegend Cat. No. 285169)

Antigen Details

Structure
C-type lectin, type II glycoprotein, 28/32 kD
Distribution

Activated T cells, B cells, NK cells, granulocytes, thymocytes, platelets, Langerhans cells

Function
Lymphocyte, monocyte, and platelet activation, NK cell killing
Cell Type
B cells, Granulocytes, Langerhans cells, NK cells, Platelets, T cells, Thymocytes, Tregs
Biology Area
Costimulatory Molecules, Immunology
Molecular Family
CD Molecules
Antigen References

1. Schlossman S, et al. Eds. 1995. Leucocyte Typing V. Oxford University Press. New York.
2. Testi R, et al. 1994. Immunol. Today 15:479.

Gene ID
NA
UniProt
View information about CD69 on UniProt.org

Related FAQs

What are Flexi-Fluors?

Flexi-Fluors are rapidly made-to-order conjugated antibodies. The technology, manufacturing processes, and specifications used to create Flexi-Fluors are the same as our regular catalog products. However, the optimal concentration and performance of each Flexi-Fluor must be determined by the customer. 

How quickly will I receive my order?

We aim to ship Flexi-Fluors within 2-3 weeks of receipt of your order. However, depending on your location, shipping times may vary. 

How are Flexi-Fluors different from regular catalog products?

Flexi-Fluors are made on demand, specifically for you. Flexi-Fluors are manufactured using the same high-quality standards, and specifications as other catalog products. For faster delivery, Flexi-Fluors are not tested by flow cytometry to determine optimal concentrations or evaluate performance. This testing needs to be performed by the customer.

How do I determine the optimal concentration for using my Flexi-Fluor? How should I titrate my antibody?

Flexi-Fluors are provided at a standard 0.2 mg/mL concentration. We recommend that you titrate your antibody to determine the optimal concentration to use for your application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL volume. We recommend that you test a range of concentrations starting from 10 µg/mL. 

 

For example, make five 1:1 serial dilutions of your 0.2 mg/mL antibody.  Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/ml) to test six concentrations - 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration. 

I can’t find the antibody-dye combination that I need. When will it be available?

We continuously update our catalog, introducing scores of new products every month. Please get in touch with our Technical Service team for an update on new products or recommendations for suitable alternatives to complete your panel. Or contact Custom Solutions to inquire about our affordable custom conjugation services. 

I need help to validate the performance of my Flexi-Fluor. Who should I contact?

Please get in touch with Technical Service for assistance. 

Can I order more than 50 μg of a Flexi-Fluor?

Yes, you can order multiple vials of the same Flexi-Fluor products.  We cannot guarantee, however, that these vials will be bottled from the same lot. For bulk single-lot orders, contact our Custom Solutions team.

What is the expiration date of my Flexi-Fluor?

Expiration dates can be found on the vial label or by using our CoA lookup tool

Other Formats

View All CD69 Reagents Request Custom Conjugation
Description Clone Applications
Purified anti-human CD69 FN50 FC,CyTOF®,IHC-F
FITC anti-human CD69 FN50 FC
PE anti-human CD69 FN50 FC
PE/Cyanine5 anti-human CD69 FN50 FC
APC anti-human CD69 FN50 FC
APC/Cyanine7 anti-human CD69 FN50 FC
PE/Cyanine7 anti-human CD69 FN50 FC
Alexa Fluor® 488 anti-human CD69 FN50 FC
Alexa Fluor® 647 anti-human CD69 FN50 FC,SB
Pacific Blue™ anti-human CD69 FN50 FC
Alexa Fluor® 700 anti-human CD69 FN50 FC
Biotin anti-human CD69 FN50 FC
PerCP/Cyanine5.5 anti-human CD69 FN50 FC
PerCP anti-human CD69 FN50 FC
Brilliant Violet 421™ anti-human CD69 FN50 FC
Brilliant Violet 785™ anti-human CD69 FN50 FC
Brilliant Violet 650™ anti-human CD69 FN50 FC
Brilliant Violet 510™ anti-human CD69 FN50 FC
Brilliant Violet 605™ anti-human CD69 FN50 FC
Purified anti-human CD69 (Maxpar® Ready) FN50 FC,CyTOF®
PE/Dazzle™ 594 anti-human CD69 FN50 FC
Brilliant Violet 711™ anti-human CD69 FN50 FC
APC/Fire™ 750 anti-human CD69 FN50 FC
TotalSeq™-A0146 anti-human CD69 FN50 PG
TotalSeq™-B0146 anti-human CD69 FN50 PG
TotalSeq™-C0146 anti-human CD69 FN50 PG
Brilliant Violet 750™ anti-human CD69 FN50 FC
KIRAVIA Blue 520™ anti-human CD69 FN50 FC
Spark NIR™ 685 anti-human CD69 Antibody FN50 FC
PE/Fire™ 640 anti-human CD69 FN50 FC
Spark YG™ 581 anti-human CD69 FN50 FC
TotalSeq™-D0146 anti-human CD69 FN50 PG
APC anti-human CD69 FN50 FC
Spark Blue™ 550 anti-human CD69 FN50 FC
PE anti-human CD69 FN50 FC
Spark Red™ 718 anti-human CD69 FN50 FC
GMP PE anti-human CD69 FN50 FC
PE/Fire™ 810 anti-human CD69 FN50 FC
PE/Fire™ 744 anti-human CD69 FN50 FC
Spark PLUS UV395™ anti-human CD69 FN50 FC
Spark Blue™ 574 anti-human CD69 (Flexi-Fluor™) FN50 FC
Go To Top Version: 1    Revision Date: 09/06/2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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